血清拉曼光谱检测在鼻咽癌早期诊断中的应用价值研究

The application value of Raman spectra for serum in early nasopharyngeal carcinoma diagnosis

背景与目的:我国南方地区鼻咽癌高发,其治疗效果与临床分期密切相关。目前早期筛查诊断的主要方法是EB病毒血清学检查,但敏感性和特异性欠佳。本研究旨在探讨血清拉曼光谱的特点,并分析其在鼻咽癌早期诊断中的价值。方法:收集2009年5月—2009年12月于本院诊治的29例经病理确诊为鼻咽癌的初诊患者(鼻咽癌组)和36例健康志愿者(健康对照组)的外周静脉血2～4 mL,经离心后获得血清样品。采用inVia型共聚焦显微拉曼光谱仪进行测量,每个样品测5个点。将同一样品所测量的5条光谱进行平均,以此代表该样品的血清拉曼光谱信号,最后将两组的拉曼光谱信号数据采用主成分分析、判别分析、ROC曲线分析等进行统计学处理。结果:两组血清的平均拉曼光谱形态和谱峰基本相似,但在约900～1 000 cm-1及1 600～1 700 cm-1波段,健康对照组的血清拉曼光谱的谱峰强度高于鼻咽癌组,而在1 200～1 400 cm-1波段,健康对照组的血清拉曼光谱的谱峰强度低于鼻咽癌组。进一步采用主成分降维处理后,发现血清光谱的第一、二、三主成分对两组血清具有较好的聚类作用。判别分析及交叉检验显示,激光拉曼光谱技术对鼻咽癌组血清的判别灵敏度为89.7%(26/29),对健康对照组的特异度为91.7%(33/36)。其ROC曲线下面积为:0.971。结论:激光拉曼光谱技术可很好地区分鼻咽癌及健康人血清,值得进一步探讨其临床应用价值。

Background and purpose：Nasopharyngeal carcinoma（NPC） highly happens in southern China,and the treatment outcome of NPC is closely related to the clinical stage.Currently,EB virus serology detection is the main method of early screening and diagnosis for NPC.But sensitivity and specificity are poor.Therefore,the aim of this study was to discuss the application value of laser Raman spectra for serum screening and early diagnosis in regards to NPC.Methods：Thirty-six cases of healthy volunteers and 29 cases of newly diagnosed NPC by pathology were selected in our hospital from May 2009 to Dec.2009.About 2-4 mL peripheral venous blood samples were taken from all subjects.All blood samples were centrifuged to remove blood cells;serum samples were obtained in the supernatant.InVia confocal Raman microscope was applied to test the samples.For each sample,we tested 5 spots for 5 Raman spectra.The average of 5 spectra in 1 sample represented the signal of Raman spectra in one case.Then,Raman spectrum of NPC patients group and healthy volunteers group were analyzed by statistical methods,including principal component analysis,discrimination analysis and receiver operating characteristic curve.Results：The average of spectra shapes and peaks for the two groups were almost the same.However,in 900-1？000 cm-1 and 1？600-1？700 cm-1 wave number range,Raman intensity of healthy volunteers blood plasma samples were greater than those of NPC patients,while in the 1？200-1？400 cm-1 wave number range,more intense in the latter.Then,multivariate analysis PCA was used to integrate the spectral information into 7 principle components.It can be visualized a good separation of the 2 groups samples according to the score of the first,the second and the third principle component.Moreover,discrimination analysis was performed on these serum spectra,the sensitivity and specificity between 2 groups were 89.7%（26/29） and 91.7%（33/36）.The integrated area under the ROC curve was 0.971.Conclusion：Technology of laser Raman spectra is able to distinguish the serum in NPC from normal well.Although the results are encouraging,we need more cases to confirm it.